Gene Therapy in the Cornea

Abstract

Several advantages are apparent in making the cornea particularly attractive for gene transfer: it has a well‐defined anatomy and is easily accessible during ambulatory visits, as well as surgical procedures. The most important, and most difficult, challenge in gene therapy is the issue of delivery. Although naked or plasmid deoxyribonucleic acid (DNA) has occasionally been applied into the corneal stroma, usually, DNA is complexed in a vector to enhance delivery into the cornea. To date, the most efficient method for delivering nucleic acid‐based treatments mainly involves viral vectors, including retrovirus, lentivirus, adeno‐associated virus and adenovirus. However, nonviral vectors (lipid‐ or polymeric‐based systems), although less efficient, are safer and its production is simpler and cheaper than viral vectors. Corneal diseases potentially treatable by gene therapy include mucopolysaccharidosis VII, herpetic stromal keratitis, corneal neovascularisation, corneal burns and corneal transplantation and graft rejection, among others.

Key Concepts

  • The cornea is an ideal target for gene therapy as it is accessible, relatively immune privileged and easily monitored owing to its transparency.
  • The most important, and most difficult, challenge in corneal gene therapy is the issue of delivery.
  • Corneal diseases potentially treatable by gene therapy include mucopolysaccharidosis VII, herpetic stromal keratitis, corneal neovascularisation, corneal burns and corneal transplantation and graft rejection, among others.
  • Gene therapy in the cornea has been mainly studied in animal models, the clinical trials in humans being scarce.

Keywords: gene therapy; cornea; viral vectors; nonviral vectors; mucopolysaccharidosis VII; herpetic stromal keratitis; corneal neovascularisation; corneal scarring; corneal transplantation

Figure 1. Structure of the cornea.
Figure 2. Representative stereomicroscopy (a) and confocal microscopy (b) images showing transgene delivery in the rabbit stroma in vivo noted 2 days after topical application of transfection solution (1 µg μL−1 plasmid in 50 nmol DDAB and 50 nmol DOPE in 100 μL lactated Ringer's solution) onto the rabbit cornea via custom delivery technique. The plasmid expresses transgene under control of CMV + chicken‐β‐actin promoter. Nuclei are stained blue with DAPI. Reproduced from Mohan et al. (2013) © Elsevier.
Figure 3. Barriers that DNA must overcome to reach the nucleus.
Figure 4. Onset and duration of AAV2/8 transgene expression. (a) Fluorescence detected by in vivo microscopy 3 days postinjection of AAV2/8. (b) At this timepoint, EGFP expression (arrow) was in the corneal epithelium (asterisk) as determined by histological epifluorescence studies. EGFP (arrows) could be detected in the stroma by 1 month postinjection by in vivo (c) and histological (d) studies. EGFP expression continued throughout the mouse cornea at 6 month postinjection in vivo (e) and by histological studies (f; montage of two overlapping photographs). EGFP expression persisted at 17 month postinjection (longest timepoint tested) as seen by in vivo (g) and histological studies (h). (i, j) Processing and analysis of the stack acquisition of the 10‐µm‐thick section shown in panel (h) using the Imaris software. Magnifications (a, c, e and g): 20×. Reproduced from Hippert et al. (2012) © PLoS One (Creative Commons Attribution (CC BY) license).
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References

Alexander JJ and Hauswirth WW (2008) Adeno‐associated viral vectors and the retina. Advances in Experimental Medicine and Biology 613: 121–128.

Apaolaza PS, Del Pozo‐Rodríguez A, Torrecilla J, et al. (2015) Journal of Controlled Release 217: 273–283.

Bakunowicz‐Łazarczyk A and Urban B (2016) Assessment of therapeutic options for reducing alkali burn‐induced corneal neovascularization and inflammation. Advances in Medical Sciences 61: 101–112.

Chen P, Yin H, Wang Y, et al. (2010) Multi‐gene targeted antiangiogenic therapies for experimental corneal neovascularization. Molecular Vision 16: 310–319.

Chira S, Jackson CS, Oprea I, et al. (2015) Progresses towards safe and efficient gene therapy vectors. Oncotarget 6: 30675–30703.

Daheshia M, Kuklin N, Manickan E, et al. (1998) Immune induction and modulation by topical ocular administration of plasmid DNA encoding antigens and cytokines. Vaccine 16: 1103–1110.

Delgado D, del Pozo‐Rodríguez A, Solinís MÁ, et al. (2011) Understanding the mechanism of protamine in solid lipid nanoparticle‐based lipofection: the importance of the entry pathway. European Journal of Pharmaceutics and Biopharmaceutics 79: 495–502.

Delgado D, del Pozo‐Rodríguez A, Solinís MÁ, et al. (2012) Dextran and protamine‐based solid lipid nanoparticles as potential vectors for the treatment of X‐linked juvenile retinoschisis. Human Gene Therapy 23: 345–355.

Delgado D, del Pozo‐Rodríguez A, Angeles Solinís M, et al. (2013) New gene delivery system based on oligochitosan and solid lipid nanoparticles: ‘in vitro’ and ‘in vivo’ evaluation. European Journal of Pharmaceutical Sciences 50: 484–491.

Elbadawy HM, Gailledrat M, Desseaux C, et al. (2012) Targeting herpetic keratitis by gene therapy. Journal of Ophthalmology 2012: article ID 594869.

Elbadawy HM, Gailledrat M, Desseaux C, et al. (2014) Gene transfer of integration defective anti‐HSV‐1 meganuclease to human corneas ex vivo. Gene Ther 21: 272–281. http://www.ncbi.nlm.nih.gov/pubmed/24430237

EMA (2015) Guideline on the quality, non‐clinical and clinical aspects of gene therapy medicinal products. Draft. EMA/CAT/80183/2014. European Medicines Agency.

Foged C (2012) siRNA delivery with lipid‐based systems: promises and pitfalls. Current Topics in Medicinal Chemistry 12: 97–107.

de la Fuente M, Seijo B and Alonso MJ (2008) Novel hyaluronic acid‐chitosan nanoparticles for ocular gene therapy. Investigative Ophthalmology & Visual Science 49: 2016–2024.

Gong N, Pleyer U, Vogt K, et al. (2007) Local overexpression of nerve growth factor in rat corneal transplants improves allograft survival. Investigative Ophthalmology & Visual Science 48: 1043–1052.

Hacein‐Bey‐Abina S, Von Kalle C, Schmidt M, et al. (2003) LMO2‐associated clonal T cell proliferation in two patients after gene therapy for SCID‐X1. Science 302: 415–419.

Hacein‐Bey‐Abina S, Garrigue A, Wang GP, et al. (2008) Insertional oncogenesis in 4 patients after retrovirus‐mediated gene therapy of SCID‐X1. Journal of Clinical Investigation 118: 3132–3142.

Hattori M, Shimizu K and Katsumura K (2012) Effects of all‐trans retinoic acid nanoparticles on corneal epithelial wound healing. Graefe's Archive for Clinical and Experimental Ophthalmology 250: 557e563.

Hippert C, Ibanes S, Serratrice N, et al. (2012) Corneal transduction by intra‐stromal injection of AAV vectors in vivo in the mouse and ex vivo in human explants. PLoS One 7: e35318.

Jani PD, Singh N, Jenkins C, et al. (2007) Nanoparticles sustain expression of Flt intraceptors in the cornea and inhibit injury‐induced corneal angiogenesis. Investigative Ophthalmology & Visual Science 48: 2030–2036.

Kamata Y, Okuyama T, Kosuga M, et al. (2001) Adenovirus‐mediated gene therapy for corneal clouding in mice with mucopolysaccharidosis type VII. Molecular Therapy: The Journal of the American Society of Gene Therapy 4: 307–312.

Kampik D, Ali RR and Larkin DF (2012) Experimental gene transfer to the corneal endothelium. Experimental Eye Research 95: 54–59.

Klausner EA, Peer D, Chapman RL, et al. (2007) Corneal gene therapy. Journal of Controlled Release 124: 107–133.

Klausner EA, Zhang Z, Chapman RL, et al. (2010) Ultrapure chitosan oligomers as carriers for corneal gene transfer. Biomaterials 31: 1814–1820.

Klebe S, Sykes PJ, Coster DJ, et al. (2001) Prolongation of sheep corneal allograft survival by ex vivo transfer of the gene encoding interleukin‐10. Transplantation 71: 1214–1220.

Ljubimov AV and Saghizadeh M (2015) Progress in corneal wound healing. Progress in Retinal and Eye Research 49: 17–45.

Mohan RR, Rodier JT and Sharma A (2013) Corneal gene therapy: basic science and translational perspective. The Ocular Surface 11 (3): 150–164.

Mashhour B, Couton D, Perricaudet M, et al (1994) In vivo adenovirusmediated gene transfer into ocular tissues. Gene Therapy 1: 122–126.

Masuda I, Matsuo T, Yasuda T, et al. (1996) Gene transfer with liposomes to the intraocular tissues by different routes of administration. Investigative Ophthalmology & Visual Science 37: 1914–1920.

Muenzer J (2011) Overview of the mucopolysaccharidoses. Rheumatology 50 (Suppl 5): v4–v12.

Nosov M, Wilk M, Morcos M, et al. (2012) Role of lentivirus‐mediated overexpression of programmed death‐ligand 1 on corneal allograft survival. American Journal of Transplantation 12: 1313–1322.

Oliveira C, Silveira I, Veiga F, et al. (2015) Recent advances in characterization of nonviral vectors for delivery of nucleic acids: impact on their biological performance. Expert Opinion on Drug Delivery 12: 27–39.

Parker DG, Kaufmann C, Brereton HM, et al. (2007) Lentivirus‐mediated gene transfer to the rat, ovine and human cornea. Gene Therapy 14: 760–767.

Parker DG, Coster DJ, Brereton HM, et al. (2010) Lentivirus‐mediated gene transfer of interleukin 10 to the ovine and human cornea. Clinical & Experimental Ophthalmology 38: 405–413.

Pleyer U and Schlickeiser S (2009) The taming of the shrew? The immunology of corneal transplantation. Acta Ophthalmologica 87: 488–497.

del Pozo‐Rodríguez A, Pujals S, Delgado D, et al. (2009) A proline‐rich peptide improves cell transfection of solid lipid nanoparticle‐based non‐viral vectors. Journal of Controlled Release 133: 52–59.

Qazi Y and Hamrah P (2013) Gene therapy in corneal transplantation. Seminars in Ophthalmology 28: 287–300.

Rivera VM, Gao GP, Grant RL, et al. (2005) Long‐term pharmacologically regulated expression of erythropoietin in primates following AAV‐mediated gene transfer. Blood 105: 424–1430.

Ritter T and Pleyer U (2009) Novel gene therapeutic strategies for the induction of tolerance in cornea transplantation. Expert Review of Clinical Immunology 5: 749–764.

Ritter T, Wilk M and Nosov M (2013) Gene therapy approaches to prevent corneal graft rejection: where do we stand? Ophthalmic Research 50: 135–140.

Rodríguez‐Gascón A, Del Pozo‐Rodríguez A, Isla A, et al. (2015) Vaginal gene therapy. Advanced Drug Delivery Reviews 92: 71–83.

Serratrice N, Cubizolle A, Ibanes S, et al. (2014) Corrective GUSB transfer to the canine mucopolysaccharidosis VII cornea using a helper‐dependent canine adenovirus vector. Journal of Controlled Release 181: 22–31.

Solinís MÁ, del Pozo‐Rodríguez A, Apaolaza P, et al. (2015) Treatment of ocular disorders by gene therapy. European Journal of Pharmaceutics and Biopharmaceutics 95 (Pt B): 331–342.

Somia N and Verma IM (2000) Gene therapy: trials and tribulations. Nature Reviews Genetics 1: 91–99.

Stechschulte SU, Joussen AM, von Recum HA, et al. (2001) Rapid ocular angiogenic control via naked DNA delivery to cornea. Investigative Ophthalmology & Visual Science 42: 1975–1979.

Testa F, Maguire AM, Rossi S, et al. (2013) Three‐year follow‐up after unilateral subretinal delivery of adeno‐associated virus in patients with Leber congenital Amaurosis type 2. Ophthalmology 120: 1283–12891.

Willett K and Bennett J (2013) Immunology of AAV‐mediated gene transfer in the eye. Frontiers in Immunology 4: 261.

Williams KA and Coster DJ (2010) Gene therapy for diseases of the cornea ‐ a review. Clinical & Experimental Ophthalmology 38: 93–103.

Yu H, Wu J, Li H, et al. (2007) Inhibition of corneal neovascularization by recombinant adenovirus‐mediated sFlk‐1 expression. Biochemical and Biophysical Research Communications 361: 946–952.

Further Reading

Collins M and Thrasher A (2015) Gene therapy: progress and predictions. Proceedings of the Royal Society B 282: pii: 20143003.

Kotterman MA, Chalberg TW and Schaffer DV (2015) Viral vectors for gene therapy: translational and clinical outlook. Annual Review of Biomedical Engineering 17: 63–89.

Meek KM and Knupp C (2015) Corneal structure and transparency. Progress in Retinal and Eye Research 49: 1–16.

Rodríguez‐Gascón A, del Pozo‐Rodríguez A and Solinís MA (2012) Non‐viral delivery systems in gene therapy. In: Molina FM (ed) Gene Therapy ‐ Tools and Potential Applications, pp. 3–33, chap. 1. Rijeka, Croatia: Intech.

Sharma A, Ghosh A, Siddappa C, et al. (2010) Ocular surface: gene therapy. In: Besharse J, Dana R and Dartt DA (eds) Encyclopedia of the eye, pp. 185–194. Atlanta: Elsevier Inc.

Wang Y, Rajala A and Rajala RV (2015) Lipid nanoparticles for ocular gene delivery. Journal of Functional Biomaterials 6: 379–394.

Yellepeddi VK and Palakurthi S (2015) Recent advances in topical ocular drug delivery. Journal of Ocular Pharmacology and Therapeutics 32: 67–82.

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Rodríguez‐Gascón, Alicia, del Pozo‐Rodríguez, Ana, Isla, Arantxaxu, and Solinís, María A(Jun 2016) Gene Therapy in the Cornea. In: eLS. John Wiley & Sons Ltd, Chichester. http://www.els.net [doi: 10.1002/9780470015902.a0024274]